BioLife Solutions Inc. is a Biological Packaging Company dedicated to developing improved solutions for hypothermic (4 to 8C) and cryopreservation (-196C) of human cells and tissues. The Company generates revenues from direct sales of its two platform solutions, HypoThermosol (HTS) and CryoStor as well as executing Custom Solution Optimization Program contracts with cell therapy and tissue engineering companies. The HTS platform consists of HTS-BASE (the original HTS formulation); HTS-DCC (a formulation that controls cation flux); HTS-FRS (a formulation that controls free radical accumulation) and HTS-LIVER (a formulation that controls apoptosis in hepatocytes). The first three HTS formulations are available through MediaTech Inc, whereas the fourth is currently under development. CryoStor is an HTS solution designed for improved cryopreservation necessitating reduced amounts of cryoprotective agent (e.g. DMSO). BioLife has also launched a program in Packaging and Downloadable Diagnostics that will allow cell therapy/tissue engineering companies who currently use HTS to track the history of temperature, shock, and pressure changes that occur during cell product transit. BioLife's HTS platform solutions were designed originally to protect substrate-attachment cells. But the recent clinical success of HTS-FRS preservation of suspended human myoblasts used in BioHeart Inc's successful European cellular cardiomyoplasty trials, coupled with CryoStor's apparent superiority in cryopreserving ATCC human hybridomas, experiments recently performed by ATCC personnel, has demanded that BioLife launch a new research program to understand the stress mechanisms that occur to suspended cells prior to preservation. Four Specific Aims are proposed as follows: SAI: Improving preservation by occupying integrin receptors that, in turn, could block anoikis; SA2: Improving preservation through inhibiting cell shape change that occurs due to the suspension process; SA3: Improving preservation by blocking FAS activation, a known death receptor that the Company has linked to preservation efficacy; SA4: Test the dual-function free radical inhibitor, Emblica, to determine if a combination free radical/metal chelator can improve the preservation of suspended cells. The intent of this Phase I SBIR project is to better understand the stress mechanisms that need to be controlled for improved preservation of suspended cells that will lead to the Phase II development of HTS-SUSP.